Imlay J A
Department of Microbiology, University of Illinois, Urbana 61801, USA.
J Biol Chem. 1995 Aug 25;270(34):19767-77.
Aerobic organisms synthesize superoxide dismutases in order to escape injury from endogenous superoxide. An earlier study of Escherichia coli indicated that intracellular superoxide is formed primarily by autoxidation of components of the respiratory chain. In order to identify those components, inverted respiratory vesicles were incubated with five respiratory substrates. In most cases, essentially all of the superoxide was formed through autoxidation of fumarate reductase, despite the paucity of this anaerobic terminal oxidase in the aerobic cells from which the vesicles were prepared. In contrast, most dehydrogenases, the respiratory quinones, and the cytochrome oxidases did not produce any detectable superoxide. The propensity of fumarate reductase to generate superoxide could conceivably deluge cells with superoxide when anaerobic cells, which contain abundant fumarate reductase, enter an aerobic habitat. In fact, deletion or overexpression of the frd structural genes improved and retarded, respectively, the outgrowth of superoxide dismutase-attenuated cells when they were abruptly aerated, suggesting that fumarate reductase is a major source of superoxide in vivo. Steric inhibitors that bind adjacent to the flavin completely blocked superoxide production, indicating that the flavin, rather than an iron-sulfur cluster, is the direct electron donor to oxygen. Since the turnover numbers for superoxide formation by other flavoenzymes are orders of magnitude lower than that of fumarate reductase (1600 min-1), additional steric or electronic factors must accelerate its autoxidation.
需氧生物合成超氧化物歧化酶以避免内源性超氧化物造成的损伤。早期对大肠杆菌的研究表明,细胞内超氧化物主要由呼吸链成分的自氧化形成。为了确定这些成分,将反向呼吸囊泡与五种呼吸底物一起孵育。在大多数情况下,基本上所有的超氧化物都是通过富马酸还原酶的自氧化形成的,尽管在制备囊泡的需氧细胞中这种厌氧末端氧化酶含量很少。相比之下,大多数脱氢酶、呼吸醌和细胞色素氧化酶都不会产生任何可检测到的超氧化物。当含有丰富富马酸还原酶的厌氧细胞进入需氧环境时,富马酸还原酶产生超氧化物的倾向可能会使细胞被超氧化物淹没。事实上,当超氧化物歧化酶减弱的细胞突然通气时,frd结构基因的缺失或过表达分别改善和延缓了它们的生长,这表明富马酸还原酶是体内超氧化物的主要来源。与黄素相邻结合的空间抑制剂完全阻断了超氧化物的产生,这表明黄素而不是铁硫簇是氧的直接电子供体。由于其他黄素酶形成超氧化物的周转数比富马酸还原酶(1600分钟-1)低几个数量级,额外的空间或电子因素必定加速了它的自氧化。